该研究以透明质酸钠(sodium hyaluronate, SH)、乳清蛋白(whey protein, WP)复合物为外水相W2,采用单因素试验和星点设计效应面法(central composite design response surface method,CCD-RSM)构建了负载亲脂性二氢槲皮素(dihydroquercetin, DHQ)和亲水性表没食子儿茶素没食子酸酯[(-)-epigallocatechin gallate, EGCG]的水包油包水(water-in-oil-in-water, W1/O/W2)双重乳液共递送体系。以聚甘油蓖麻醇酸酯(polyglyceryl-6 polyricinoleate, PGPR)体积分数和水油体积比为影响因素,通过乳液表、微观形态及粒径筛选出制备W1/O初乳的最佳配比:PGPR体积分数4%、初乳水油体积比20∶1。通过单因素试验筛选外水相W2(SH-WP)质量比、初乳体积比、分散转数及时间,在此基础上采用CCD-RSM考察W2质量比、初乳体积比对W1/O/W2双乳粒径、zeta电位影响,用二项式及多元线性回归模型拟合建立指标与因素之间的关系,确定最优工艺参数:SH-WP质量比3∶1、初乳体积比为63.32%、分散转速12 000 r/min、分散时间3 min,优化工艺各指标的预测值和目标值接近。制备的乳液表观白皙、体系均一稳定,共聚焦激光扫描显微镜(confocal laser scanning microscope, CLSM)染色图像表明该乳为典型的“两膜三相”界面结构。通过傅里叶红外光谱、圆二色谱、荧光光谱对SH-WP复合物的结构探究表明,SH和WP主要为静电吸附和氢键相互作用;三相接触角探究表明,SH-WP复合物的表面润湿性得到明显改善。所制备的双重乳液对EGCG、DHQ具有良好的负载能力,包埋率分别为41.24%、95.34%。综上表明,该研究制备的W1/O/W2双重乳液可同时实现亲水性EGCG、亲脂性DHQ的共同递送,具有良好的活性成分递送潜力。
The complex of sodium hyaluronate (SH) and whey protein (WP) was used as the external aqueous phase W2.A water-in-oil-in-water (W1/O/W2) double emulsion co-delivery system loaded with lipophilic dihydroquercetin (DHQ) and hydrophilic (-)-epigallocatechin gallate (EGCG) was constructed by single factor test and central composite design response surface method.Taking the concentration of polyglyceryl-6 polyricinoleate (PGPR) and the volume ratio of water to oil as the influencing factors, the optimum ratio of W1/O colostrum was screened by emulsion surface, microstructure and particle size:the concentration of PGPR was 4% (v/v), and the volume ratio of water to oil was 20% (v/v).Through single factor experiments, the mass ratio of W2 (SH-WP) in the external aqueous phase, the volume ratio of colostrum, the dispersion speed and time were screened.Based on this, the star point design effect surface method (CCD-RSM) was used to investigate the effects of W2 mass ratio and colostrum volume ratio on the particle size and Zeta potential of W1/O/W2 emulsions.The binomial and multiple linear regression models were used to fit and establish the relationship between indicators and factors, and the optimal process parameters were determined:SH-WP mass ratio of 3∶1, colostrum volume ratio of 63.32%, dispersion speed of 12 000 r/min, and dispersion time of 3 min.The predicted values of various indicators in the optimized process were close to the target values.The prepared emulsion has a white appearance and a uniform and stable system.The confocal laser scanning microscope (CLSM) staining image shows that the emulsion is a typical “two-film three-phase” interface structure.The structure of SH-WP complex was investigated by Fourier transform infrared spectroscopy, circular dichroism and fluorescence spectroscopy.The results showed that SH and WP were mainly electrostatic adsorption and hydrogen bond interaction.The three-phase contact angle investigation showed that the surface wettability of the SH-WP composite was significantly improved.The embedding rates of EGCG and DHQ in the prepared double emulsion were 41.24% and 95.34%, respectively, and the double emulsion had good stability.In summary, the W1/O/W2 double emulsion prepared in this study can simultaneously achieve the co-delivery of hydrophilic EGCG and lipophilic DHQ, and has good potential for active ingredient delivery.
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